Table 4.

Rough Limit of Detection (LoD) for each RT-ddPCR reaction. Dilutions were measured in quadruplicate in RT-ddPCR and in duplicate in RT-PCR (using primer/probe set from [43] as described in [44]). Targets with mismatches to both FAM and VIC probes are indicated with blue type. Mean concentrations are those measured in RT-ddPCR replicates at the LoD; for the dilution beyond LoD (gray type), concentrations are calculated from the LoD.

Reaction	Amino acid(s)	E Ct	Mean copies/µL	Mean copies/Rxn	Positive replicates
417	K	33.4	2.22	22.2	4/4	At LoD
	N	35.7	0.77	7.7	4/4
	T	35.1	1.39	13.9	4/4
	K	37.1	0.22	2.2	1/4	Beyond LoD
	N	37.3	0.07	0.8	0/4
	T	NDET	0.14	1.4	0/4
484	E	33.4	0.65	6.5	4/4	At LoD
	K	35.1	0.88	8.8	4/4
	Q	30.8	13.10	131.0	4/4
	E	37.1	0.07	0.7	0/4	Beyond LoD
	K	NDET	0.09	0.9	0/4
	Q	34.1	1.31	13.1	3/4
452/501	L/N	33.4	1.17	11.7	4/4	At LoD
	L/Y	35.1	0.86	8.6	4/4
	R/N	34.1	0.91	9.1	4/4
	L/N	37.1	0.11	1.2	1/4	Beyond LoD
	L/Y	NDET	0.09	0.9	0/4
	R/N	NDET	0.09	0.9	2/4